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Verocytotoxin expression in Escherichia coli
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Fox, Sian L. (1992) Verocytotoxin expression in Escherichia coli. PhD thesis, University of Warwick.
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WRAP_Theses_Fox_1992.pdf - Submitted Version - Requires a PDF viewer. Download (6Mb) | Preview |
Official URL: http://webcat.warwick.ac.uk/record=b3252313~S15
Abstract
In order to examine the physiology of verotoxin (VT) production in vitro, it was necessary to develop an assay to enable specific measurement of VTl and VT2 expression. Initial efforts focussed on quantifying VTl and VT2 messenger RNA synthesis. RNA extraction and electrophoresis was consistently achieved, however, subsequent probing of Northern filters was not successful.
A gene fusion between the slt-ll operon and the transposon vector TnphoA, such that expression of the phoA gene was brought under the control of the 5/MI promoter, was then developed. The presence and site of insertion of TnphoA within the s/MI operon was confirmed by restriction and sequence analysis, and a single copy fusion derivative within the VT2-producing E. coli strain E32511 was then obtained by exchange recombination. Use of the resulting single copy fusion derivative (E32511 SLF22/1) and a plasmid encoded slt-\. fïnphoA fusion (pSC105) demonstrated significant differences in the synthesis, secretion and localisation abilities of VTl and VT2 during growth under aerobic, anaerobic and iron limiting conditions. These characteristics could have important implications on the relative abilities of the two toxins to cause disease in vivo.
| Item Type: | Thesis (PhD) | ||||
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| Subjects: | Q Science > QP Physiology Q Science > QR Microbiology |
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| Library of Congress Subject Headings (LCSH): | Verocytotoxins, Escherichia coli | ||||
| Official Date: | September 1992 | ||||
| Dates: |
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| Institution: | University of Warwick | ||||
| Theses Department: | Department of Biological Sciences | ||||
| Thesis Type: | PhD | ||||
| Publication Status: | Unpublished | ||||
| Supervisor(s)/Advisor: | Dow, Crawford S. | ||||
| Format of File: | |||||
| Extent: | xxiii, 265 leaves : illustrations | ||||
| Language: | eng |
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